發(fā)布時(shí)間：2018-06-23 05:36

  本文選題：野生動物 + 腸道微生物��； 參考：《吉林農(nóng)業(yè)大學(xué)》2015年碩士論文

【摘要】：目前,國內(nèi)外研究人員利用現(xiàn)代分子生物學(xué)技術(shù)在動物腸道微生物研究領(lǐng)域取得諸多進(jìn)展,如確立了宿主與微生物之間的相互關(guān)系,提高了宿主的健康狀態(tài)和營養(yǎng)水平,增加了動物對飼料的利用效率,明確了胃腸道微生物的代謝通路等。近年來對野生動物腸道微生態(tài)的研究得到廣泛關(guān)注,研究野生動物腸道微生物不僅可以描述野生動物腸道微生物的系統(tǒng)發(fā)育,分析其腸道菌群功能,研究其胃腸道微生物區(qū)系變化規(guī)律、信號通路及其響應(yīng)機(jī)制,而且可以通過改變腸道微生物組成、增加優(yōu)勢菌群數(shù)量等有效措施預(yù)防和治療某些腸道疾病,進(jìn)而為野生動物健康可持續(xù)的繁殖和生存做出貢獻(xiàn)。本研究采用PCR-DGGE技術(shù)研究不同野生動物腸道菌群多樣性,經(jīng)分析得到23條差異條帶,測序結(jié)果表明,厚壁菌門(Firmicutes)為腸道優(yōu)勢菌群。對草食動物與肉食動物腸道菌群分析可知,芽孢桿菌科(Bacillaceae)的梭菌屬(Fusobacterium)在草食動物腸道菌群的屬水平占優(yōu)勢地位。在反芻動物糞便中檢測出具有高效纖維分解能力的溶纖維丁酸弧菌(Butyrivibrio fibrisolvens)。消化鏈球菌屬(Peptostreptococcus)為肉食動物腸道菌群的優(yōu)勢菌屬,在犬科肉食動物糞便中檢測到Anaerovorax,在貓科動物糞便中分離出特有菌為河流漫游球菌(Vagococcus fluvialis)。本研究同時(shí)選擇宏基因組測序技術(shù)研究野生動物腸道菌群多樣性,共得到242 609條序列,40 130個(gè)操作分類單元(OTUs),其中在草食動物腸道菌群中占優(yōu)勢地位的細(xì)菌歸類為Firmicutes、擬桿菌門(Bacteroidetes)、變形菌門(Proteobacteria)、放線菌門(Actinobacteria)和疣微菌門(Verrucomicrobia),肉食動物腸道菌群中優(yōu)勢菌門為Firmicutes、Actinobacteria、Proteobacteria、Bacteroidetes和梭桿菌門(Fusobacteria),在屬水平上,草食動物腸道微生物明顯比肉食動物腸道微生物種類豐富,在單胃草食動物和反芻動物腸道菌群比較中,具有明顯差異的菌屬為瘤胃球菌屬(Ruminococcus)、顫桿菌克(Oscillibacter)、鳥桿菌屬(Ornithobacterium)、Anaerovorax、Paludibacter、Alistipes和Succinivibrio。在肉食動物中,犬科動物和貓科動物腸道微生物多樣性表現(xiàn)出差異的菌屬為醋弧菌屬(Acetivibrio)、假單胞菌屬(Pseudomonas)、Sporobacter、Bulleidia和Eggerthella。在現(xiàn)代微生物多樣性研究方法基礎(chǔ)上,本研究進(jìn)一步利用傳統(tǒng)分離培養(yǎng)方法對野生動物糞便中菌群進(jìn)行分離、鑒定,然后對菌株的纖維素酶、木質(zhì)素酶、蛋白酶、淀粉酶及脂肪酶相關(guān)功能進(jìn)行研究,同時(shí)根據(jù)菌株功能對腸道菌群功能進(jìn)行分析,并對PCR-DGGE技術(shù)和宏基因組測序技術(shù)所得結(jié)果進(jìn)行驗(yàn)證,發(fā)現(xiàn)利用宏基因組測序技術(shù)得到的結(jié)果較PCR-DGGE技術(shù)更加全面;并確定草食動物與肉食動物糞便中存在相同菌株,它們是Bacillus drentensis和Bacillus safensis,發(fā)現(xiàn)其具有產(chǎn)纖維素酶、蛋白酶和淀粉酶功能;分析菌群多樣性與菌株功能之間關(guān)系時(shí)發(fā)現(xiàn),從不同食性動物糞便中分離出的菌株在功能上有所差異,從草食動物糞便中分離出的菌株多具有纖維素分解功能,而從肉食動物糞便中分離的菌株多具有脂肪分解功能。通過以上研究,所得結(jié)果能夠初步反映不同野生動物腸道菌群結(jié)構(gòu)及其功能特征,這為今后野生動物腸道菌群多樣性的研究以及功能優(yōu)勢菌株的篩選提供一定理論依據(jù),并為野生動物消化功能相關(guān)基因研究以及野生動物健康繁殖與生存奠定一定基礎(chǔ)。
[Abstract]:At present, researchers at home and abroad have made great progress in the field of animal intestinal microflora by modern molecular biology technology, such as establishing the relationship between host and microorganism, improving the health and nutrition level of the host, increasing the utilization efficiency of animal feed and clarified the metabolic pathways of gastrointestinal microorganism. In recent years, the research on the intestinal microflora of wild animals has been widely concerned. The study on the intestinal microorganism of wild animals can not only describe the phylogenetic development of the intestinal microbes in wild animals, analyze the intestinal microflora function, study the changes in the microbiological flora of the gastrointestinal tract, the signaling pathway and its response mechanism, and can also change the intestinal tract by changing the intestinal tract. Microbial composition, increasing the number of dominant bacteria groups, and other effective measures to prevent and treat certain intestinal diseases, and thus contribute to the healthy and sustainable reproduction and survival of the wild animals. This study used PCR-DGGE technology to study the diversity of intestinal flora in different wild animals. 23 different bands were analyzed, and the results of sequencing showed that Fir Micutes) for the intestinal predominant flora. The analysis of the intestinal flora of herbivorous and carnivorous animals showed that the Fusobacterium of Bacillaceae was dominant in the level of intestinal flora of herbivorous animals. In ruminant feces, the soluble Vibrio butyrate (Butyrivibrio FIB with high efficiency of fibrinolysis) was detected. Risolvens. Peptostreptococcus is the dominant bacteria in the intestinal flora of carnivorous animals. Anaerovorax is detected in the faeces of canine carnivorous animals, and the endemic bacteria isolated from the faeces of the cat family is Vagococcus fluvialis (Vagococcus fluvialis). This study selected the macrogenome sequencing technology to study the intestines of wild animals in the same time. There were 242609 sequences and 40130 operating taxa (OTUs), among which the dominant bacteria in the herbivorous intestinal flora were classified as Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria and Verrucomicrobia, and the intestinal flora of carnivorous animals The dominant bacteria were Firmicutes, Actinobacteria, Proteobacteria, Bacteroidetes and Clostridium (Fusobacteria). At the level of the genus, the intestinal microbes of the herbivorous animals were obviously more abundant than those of the carnivorous animals. In the comparison of the intestinal flora of the single stomach herbivore and the ruminant, the bacteria with the obvious difference were the rumen coccus (R). Uminococcus), Oscillibacter, Ornithobacterium, Anaerovorax, Paludibacter, Alistipes and Succinivibrio. in carnivorous animals. The diversity of intestinal microbial diversity in canine and feline intestinal microflora is a genus of Vibrio (Acetivibrio), Pseudomonas (Pseudomonas), Sporobacter, Bulleidia and Eggerthella. on the basis of modern microbial diversity research methods, this study further uses the traditional isolation and culture methods to isolate and identify the bacteria in the faeces of wild animals, and then studies the related work of cellulase, lignin, protease, amylase and lipase, and the function of the strains to the intestinal flora. The function is analyzed, and the results obtained by PCR-DGGE technology and macrogenome sequencing technology are verified. It is found that the results obtained by the macro genome sequencing technology are more comprehensive than the PCR-DGGE technology, and determine the same strains in the faeces of herbivorous animals and carnivorous animals, they are Bacillus drentensis and Bacillus safensis. With the function of producing cellulase, protease and amylase, the analysis of the relationship between the diversity of the flora and the function of the strain found that the strains isolated from the faeces of different food animals were different in function, and most of the strains isolated from the faeces of herbivorous faeces had the power of cellulose decomposition, and the strains isolated from the faeces of carnivorous animals were many. The results can preliminarily reflect the structure and functional characteristics of intestinal microflora in different wild animals. This provides a theoretical basis for the study of the diversity of intestinal flora and the screening of functional dominant strains in the future, and the study of the genes related to the digestive function of wild animals and the wild animals. It lays a foundation for the healthy reproduction and survival of animals.
【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S852.6

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 楊曼;蘭阿峰;郭素芬;丁小維;鄧百萬;;免培養(yǎng)法研究野生川金絲猴腸道內(nèi)生細(xì)菌多樣性[J];微生物學(xué)通報(bào);2014年08期

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本文編號：2056022